A study with model substrates of the structure of the sites phosphorylated by rat liver casein kinase TS.
Two new sites phosphorylated by rat liver cyclic AMP-independent casein kinase TS have been identified in denatured pepsin and soybean antiprotease C-II, exhibiting the sequences: Cys-Ser-Ser(P)-Ile-Asp-Ser and His-Ser3(P)-Asp-Asp-Glu, respectively. Their phosphorylation efficiency has been compared to that of previously identified sites and the effects of chemical modifications in the vicinity of the phosphorylatable residue have been studied. The results obtained support the following conclusions: 1. All sites affected by casein kinase TS conform to the sequence: Ser/Thr-X-Glu/Asp which is also believed to be required by the mammary gland casein kinase. Threonine appears to be less suitable for phosphorylation then serine. The presence of some additional residues on the C-terminal side also appears to be required. 2. X can be either an additional acidic residue or a natural one, but not a basic residue. The contiguity of an acidic cluster to the C-terminal side of the target greatly improves the phosphorylation efficiency. 3. The residues N-terminal to the target one do not seem to be relevant for determining the site recognition by the protein kinase. 4. The predicted secondary structure constantly occurring at the phosphorylation sites is the beta-turn: apparently the bend must include both the target residue and the acidic determinant at the n + 2 position.
D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D047388:Casein Kinases; D002850:Chromatography, Gel; D003488:Cyanogen Bromide; D007700:Kinetics; D008099:Liver / Q000201:enzymology; D010434:Pepsin A / Q000378:metabolism; D010766:Phosphorylation; D010940:Plant Proteins; D011480:Protease Inhibitors / Q000378:metabolism; D011494:Protein Kinases / Q000378:metabolism*; D051381:Rats
D010940:Plant Proteins; D011480:Protease Inhibitors; C032845:antiprotease C-II; D011494:Protein Kinases; D047388:Casein Kinases; D010434:Pepsin A; D003488:Cyanogen Bromide